Bioprocess and Biosystems Engineering最新文献

The probiotic fermentation of the bioactive substance gamma-aminobutyric acid (GABA) is an attractive research topic. There is still room for further improvement in reported GABA fermentation methods based on a single substrate (l-glutamic acid or l-monosodium glutamate). Here, we devised a pH auto-buffering strategy to facilitate the fermentation of GABA by Levilactobacillus brevis CD0817. This strategy features a mixture of neutral monosodium l-glutamate plus acidic l-glutamic acid as the substrate. This mixture provides a mild initial pH; moreover, the newly dissolved l-glutamic acid automatically offsets the pH increase caused by substrate decarboxylation, maintaining the acidity essential for GABA fermentation. In this study, a flask trial was first performed to optimize the GABA fermentation parameters of Levilactobacillus brevis CD0817. The optimized parameters were further validated in a 10 L fermenter. The flask trial results revealed that the appropriate fermentation medium was composed of powdery l-glutamic acid (750 g/L), monosodium l-glutamate (34 g/L [0.2 mol/L]), glucose (5 g/L), yeast extract (35 g/L), MnSO₄·H₂O (50 mg/L [0.3 mmol/L]), and Tween 80 (1.0 g/L). The appropriate fermentation temperature was 30 °C. The fermenter trial results revealed that GABA was slowly synthesized from 0–4 h, rapidly synthesized until 32 h, and finally reached 353.1 ± 8.3 g/L at 48 h, with the pH increasing from the initial value of 4.56 to the ultimate value of 6.10. The proposed pH auto-buffering strategy may be popular for other GABA fermentations.

The production of keratinases was evaluated in submerged fermentation with Aspergillus niger and by pigs’ swine hair in a batch bioreactor. Experimental planning was performed to assess the interaction between different variables. The enzyme extract produced was characterized at various pH and temperatures and subjected to enzyme concentration using a biphasic aqueous system and salt/solvent precipitation techniques. In addition, the substrate’s potential in reducing hexavalent chromium from synthetic potassium dichromate effluent with an initial concentration of 20 mg L⁻¹ of chromium was evaluated. The resulting enzyme extract showed 89 ± 2 U mL⁻¹ of keratinase. The enzyme concentration resulted in a purification factor of 1.3, while sodium chloride/acetone and ammonium sulfate/acetone resulted in a purification factor of 1.9 and 1.4, respectively. Still using the residual substrate of swine hair from the fermentation, a 94% reduction of hexavalent chromium concentration occurred after 9 h of reaction. Thus, the study proved relevant for producing keratinases, with further environmental applicability and the possibility of concentrating the extract via low-cost processes.

Perfusion cell-culture mode has caught industrial interest in the field of biomanufacturing in recent years. Thanks to new technology, perfusion-culture processes can support higher cell densities, higher productivities and longer process times. However, due to the inherent operational complexity and high running costs, the development and design of perfusion-culture processes remain challenging. Here, we present a model-based approach to design optimized perfusion cultures of Chinese Hamster Ovary cells. Initially, four batches of bench-top reactor continuous-perfusion-culture data were used to fit the model parameters. Then, we proposed the model-based process design approach, aiming to quickly find out the “theoretically optimal” operational parameters combinations (perfusion rate and the proportion of feed medium in perfusion medium) which could achieve the target steady-state VCD while minimizing both medium cost and perfusion rate during steady state. Meanwhile, we proposed a model-based dynamic operational parameters-adjustment strategy to address the issue of cell-growth inhibition due to the high osmolality of concentrated perfusion medium. In addition, we employed a dynamic feedback control method to aid this strategy in preventing potential nutrient depletion scenarios. Finally, we test the feasibility of the model-based process design approach in both shake flask semi-perfusion culture (targeted at 5 × 10⁷ cells/ml) and bench-top reactor continuous perfusion culture (targeted at 1.1 × 10⁸ cells/ml). This approach significantly reduces the number of experiments needed for process design and development, thereby accelerating the advancement of perfusion-mode cell-culture processes.

Antibiotics are widely used as fungicides because of their antibacterial and bactericidal effects. However, it is necessary to control their dosage. If the amount of antbiotics is too much, it cannot be completely metabolized and absorbed, will pollute the environment, and have a great impact on human health. Many antibiotics usually left in factory or aquaculture wastewater pollute the environment, so it is vital to detect the content of antibiotics in wastewater. This article summarizes several common methods of antibiotic detection and pretreatment steps. The detection methods of antibiotics in wastewater mainly include immunoassay, instrumental analysis method, and sensor. Studies have shown that immunoassay can detect deficient concentrations of antibiotics, but it is affected by external factors leading to errors. The detection speed of the instrumental analysis method is fast, but the repeatability is poor, the price is high, and the operation is complicated. The sensor is a method that is currently increasingly studied, including electrochemical sensors, optical sensors, biosensors, photoelectrochemical sensors, and surface plasmon resonance sensors. It has the advantages of fast detection speed, high accuracy, and strong sensitivity. However, the reproducibility and stability of the sensor are poor. At present, there is no method that can comprehensively integrate the advantages. This paper aims to review the enrichment and detection methods of antibiotics in wastewater from 2020 to the present. It also aims to provide some ideas for future research directions in this field.

Bio-inspired zinc oxide nanoparticles are gaining immense interest due to their safety, low cost, biocompatibility, and broad biological properties. In recent years, much research has been focused on plant-based nanoparticles, mainly for their eco-friendly, facile, and non-toxic character. Hence, the current study emphasized a bottom-up synthesis of zinc oxide nanoparticles (ZnO NPs) from Psidium guajava aqueous leaf extract and evaluation of its biological properties. The structural characteristic features of biosynthesized ZnO NPs were confirmed using various analytical methods, such as UV-Vis spectroscopy, X-ray diffraction (XRD), energy-dispersive X-ray analysis (EDX), Fourier transform infrared spectroscopy (FT-IR), dynamic light scattering (DLS), Scanning electron microscopy (SEM) and high-resolution transmission electron microscopy (HR-TEM). The synthesized ZnO NPs exhibited a hydrodynamic shape with an average particle size of 11.6-80.2 nm. A significant antimicrobial efficiency with minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of 40 and 27 µg/ml for Enterococcus faecalis, followed by 30 and 40 µg/ml for Staphylococcus aureus, 20 and 30 µg/ml for Staphylococcus mutans, 30 µg/ml for Candida albicans was observed by ZnO NPs. Additionally, they showed significant breakdown of biofilms of Streptococcus mutans and Candida albicans indicating their future value in drug-resistance research. Furthermore, an excellent dose-dependent activity of antioxidant property was noticed with an IC₅₀ of 9.89 µg/ml. The antiproliferative potential of the ZnO NPs was indicated by the viability of MDA MB 231 cells, which showed a drastic decrease in response to increased concentrations of biosynthesized ZnO NPs. Thus, the present results open up vistas to explore their pharmaceutical potential for the development of targeted anticancer drugs in the future.

Biosilica (BS) and spongin (SPG) from marine sponges are highlighted for their potential to promote bone regeneration. Moreover, 3D printing is introduced as a technology for producing bone grafts with optimized porous structures, allowing for better cell attachment, proliferation, and differentiation. Thus, this study aimed to characterize the BS and BS/SPG 3D printed scaffolds and to evaluate the biological effects in vitro. The scaffolds were printed using an ink containing 4 wt.% of sodium alginate. The physicochemical characteristics of BS and BS/SPG 3D printed scaffolds were analyzed by SEM, EDS, FTIR, porosity, evaluation of mass loss, and pH measurement. For in vitro analysis, the cellular viability of the MC3T3-E1 cell lineage was assessed using the AlamarBlue^® assay and confocal microscopy, while genotoxicity and mineralization potential were evaluated through the micronucleus assay and Alizarin Red S, respectively. SEM analysis revealed spicules in BS, the fibrillar structure of SPG, and material degradation over the immersion period. FTIR indicated peaks corresponding to silicon oxide in BS samples and carbon oxide and amine in SPG samples. BS-SPG scaffolds exhibited higher porosity, while BS scaffolds displayed greater mass loss. pH measurements indicated a significant decrease induced by BS, which was mitigated by SPG over the experimental periods. In vitro studies demonstrated the biocompatibility and non-cytotoxicity of scaffold extracts. .Also, the scaffolds promoted cellular differentiation. The micronucleus test further confirmed the absence of genotoxicity. These findings suggest that 3D printed BS and BS/SPG scaffolds may possess desirable morphological and physicochemical properties, indicating in vitro biocompatibility.

Biosurfactants (BSFs) are molecules produced by microorganisms from various carbon sources, with applications in bioremediation and petroleum recovery. However, the production cost limits large-scale applications. This study optimized BSFs production by Bacillus velezensis (strain MO13) using residual glycerin as a substrate. The spherical quadratic central composite design (CCD) model was used to standardize carbon source concentration (30 g/L), temperature (34 °C), pH (7.2), stirring (239 rpm), and aeration (0.775 vvm) in a 5-L bioreactor. Maximum BSFs production reached 1527.6 mg/L of surfactins and 176.88 mg/L of iturins, a threefold increase through optimization. Microbial development, substrate consumption, concentration of BSFs, and surface tension were also evaluated on the bioprocess dynamics. Mass spectrometry Q-TOF-MS identified five surfactin and two iturin isoforms produced by B. velezensis MO13. This study demonstrates significant progress on BSF production using industrial waste as a microbial substrate, surpassing reported concentrations in the literature.

Surface enhanced Raman spectroscopy (SERS) by using gold nanoparticles (AuNPs) has gained relevance for the identification of biomolecules and some cancer cells. Searching for greener NPs synthesis alternatives, we evaluated the SERS properties of AuNPs produced by using different filamentous fungi. The AuNPs were synthesized utilizing the supernatant of Botrytis cinerea, Trichoderma atroviride, Trichoderma asperellum, Alternaria sp. and Ganoderma sessile. The AuNPs were characterized by ultraviolet-visible spectroscopy (UV-Vis) to identify its characteristic surface plasmon resonance, which was located at 545 nm (B. cinerea), 550 nm (T. atroviride), 540 nm (T. asperellum), 530 nm (Alternaria sp.), and 525 nm (G. sessile). Morphology, size and crystal structure were characterized through transmission electron microscopy (TEM); colloidal stability was assessed by Z-potential measurements. We found that, under specific incubation conditions, it was possible to obtain AuNPs with spherical and quasi-spherical shapes, which mean size range depends on the fungal species supernatant with 92.9 nm (B. cinerea), 24.7 nm (T. atroviride), 16.4 nm (T. asperellum), 9.5 nm (Alternaria sp.), and 13.6 nm (G. sessile). This, as it can be expected, has an effect on Raman amplification. A micro-Raman spectroscopy system operated at a wavelength of 532 nm was used for the evaluation of the SERS features of the AuNPs. We chose methylene blue as our target molecule since it has been widely used for such a purpose in the literature. Our results show that AuNPs synthesized with the supernatant of T. atroviride, T. asperellum and Alternaria sp. produce the stronger SERS effect, with enhancement factor (EF) of 20.9, 28.8 and 35.46, respectively. These results are promising and could serve as the base line for the development of biosensors through a facile, simple, and low-cost green alternative.

This study investigated the effect of pH on poly-γ-L-diaminobutanoic acid (γ-PAB) production by Bacillus pumilus in batch fermentation. In the natural fermentation where pH was not controlled, pH decreased from initial 7.0 to 3.0 in 18 h and γ-PAB production was 428.6 mg/L. In the pH-controlled fermentation, B. pumilus tended to proliferation at higher pH, while γ-PAB synthesis was favorable at lower pH, in which the optimal pH for γ-PAB production was 4.2, and γ-PAB yield reached 2284.5 mg/L. Adopting a pH shock strategy which lasted 9 h in the pre-fermentation phase, biomass (OD₆₀₀) and γ-PAB yield of B. pumilus were obtained as 61.3 and 2794.6 mg/L, respectively, which were 10.8% and 22.4% higher than those in batch fermentation without pH shock. Subsequent fermentation of repeated pH shocks showed that a further higher productivity could be achieved, in which the final OD₆₀₀ reached 65.1, and γ-PAB production reached as high as 3482.3 mg/L, which were increased by 6.2% and 17.1% compared with those in single pH shock, respectively. This study demonstrated that B. pumilus can synthesize more γ-PAB at suboptimal pH and provided a novel approach to regulate γ-PAB synthesis.

Hydrogel nanocatalyst composed of nickel oxide (NiO) nanoparticles embedded in PVA-alginate hydrogels were potentially explored toward the reduction of anthropogenic water pollutants. The NiO nanoparticles was accomplished via green method using waste pineapple peel extract. The formation of the nanoparticles was affirmed from different analytical techniques such as UV-Vis, FTIR, XRD, TGA, FESEM, and EDS. Spherical NiO nanoparticles were obtained having an average size of 11.5 nm. The nano NiO were then integrated into PVA-alginate hydrogel matrix forming a nanocomposite hydrogel (PVALg@ NiO). The integration of nano NiO rendered an improved thermal stability to the parent hydrogel. The PVALg@ NiO hydrogel was utilized as a catalyst in the reduction of 4-nitrophenol (4-NP), potassium hexacyanoferrate (III), rhodamine B (RhB), methyl orange (MO), and malachite green (MG) in the presence of a reducing agent, i.e., NaBH₄. Under optimized conditions, the reduction reactions were completed by 4.0 min and 3.0 min for 4-NP and potassium hexacyanoferrate (III), respectively, and the rate constant was estimated to be 1.14 min^-1 and 2.15 min^-1. The rate of reduction was found to be faster for the dyes and the respective rate constants were be 0.17 s^-1 for RhB, MG and 0.05 s^-1 for MO. The PVALg@ NiO hydrogel nanocatalyst demonstrated a recyclability of four runs without any perceptible diminution in its catalytic mettle. The efficacy of the PVALg@ NiO hydrogel nanocatalyst was further examined for the reduction of dyes in real water samples collected from different sources and the results affirm its high catalytic potential. Thus, this study paves the path for the development of a sustainable hydrogel nanocatalyst for reduction of hazardous pollutants in wastewater treatment.